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3D Localization Microscopy With Ground State Depletion (GSD 3D)
3D Localization Microscopy With Ground State Depletion (GSD 3D)
Basic Principles of the GSD Super-Resolution Technology for Displaying Highly Resolved Cellular Structures in Three DimensionsThe exact position of a fluorochrome is based on the principle of temporal separation;
This requires switching of fluorochromes between on and off states followed by a sequential readout;
Super-resolution imaging
The crucial step of dSTORM/GSD is to temporarily switch off the majority of fluorophores to allow the precise localization of single fluorophores. Super-resolution as reconstructed z-stackFluorophore in the focal plane: the image appears round; Below the focus: elongated in the vertical direction;Higher than the focus position: elongated in the horizontal direction.
Section localization scheme
Super-resolution as reconstructed z-stackCoordinates in z are assigned to a specific layer in a z-stack based on a calibration of the deformed PSF by the cylinder lens.Resolution around 70 nm can be achieved.
Evolution of resolutionAlexaFluor 488AlexaFluor 532Rhodamine 6GAlexaFluor 647Others.
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